Confocal immunofluorescent images of HeLa cells, untreated (left) or phosphatase-treated (right), labeled with Phospho-Myosin Light Chain 2 (Ser19) Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Western blot analysis of extracts from HEK293 cells stimulated with ionophore A23187 for the indicated times, using Phospho-Myosin Light Chain 2 (Thr18/Ser19) Antibody.
Western blot analysis of extracts from various cell lines using Myosin Light Chain 2 (D18E2) Rabbit mAb.
Western blot analysis of extracts from HEK293 cells stimulated with ionophore A23187 for the indicated times, using Phospho-Myosin Light Chain 2 (Ser19) Antibody.
|Phospho-Myosin Light Chain 2 (Ser19) Antibody 3671||20 µl||
||H M R Dm||18||Rabbit|
|Phospho-Myosin Light Chain 2 (Thr18/Ser19) Antibody 3674||20 µl||
|Myosin Light Chain 2 (D18E2) Rabbit mAb 8505||20 µl||
||H M R||18||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
The Myosin Light Chain 2 Antibody Sampler Kit provides an economical means to detect total, phosphorylated, and dual-phosphorylated myosin light chain 2. The kit contains enough primary and secondary antibody to perform two western blot experiments.
The antibodies in the Myosin Light Chain 2 Antibody Sampler Kit detect endogenous levels of total myosin light chain 2 (smooth muscle), myosin light chain 2 when phosphorylated at Ser19 or when dually phosphorylated at Thr18 and Ser19, respectively. The antibodies do not cross-react with the cardiac isoform of Myosin Light Chain 2.
The rabbit monoclonal antibody is produced by immunizing rabbits with a synthetic peptide corresponding to residues near the carboxy terminus of human myosin light chain 2 protein. Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser19 of human myosin light chain 2, or to residues surrounding Thr18/Ser19 of human myosin light chain 2, respectively. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.
Myosin is composed of six polypeptide chains: two identical heavy chains and two pairs of light chains. Myosin light chain 2 (MLC2), also known as myosin regulatory light chain (MRLC), RLC, or LC20, has many isoforms depending on its distribution. In smooth muscle, MLC2 is phosphorylated at Thr18 and Ser19 by myosin light chain kinase (MLCK) in a Ca2+/calmodulin-dependent manner (1). This phosphorylation is correlated with myosin ATPase activity and smooth muscle contraction (2). ROCK also phosphorylates Ser19 of smooth muscle MLC2, which regulates the assembly of stress fibers (3). Phosphorylation of smooth muscle MLC2 at Ser1/Ser2 and Ser9 by PKC and cdc2 has been reported to inhibit myosin ATPase activity (4,5). Phosphorylation by cdc2 controls the timing of cytokinesis (5). Transgenic mice lacking phosphorylation sites on the cardiac muscle isoform show morphological and functional abnormalities (6).
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